Pharmaceutical composition

ABSTRACT

The present invention provides compounds of Formula I: 
                         
along with pharmaceutical compositions containing the same, and methods of use thereof in subjects in need of treatment.

RELATED APPLICATIONS

This application is a continuation of PCT/US11/51163, filed Sep. 12,2011, which application claims the benefit of U.S. Provisional PatentApplication No. 61/384,781, filed Sep. 21, 2010, the disclosures of eachof which are incorporated by reference herein in their entireties.

BACKGROUND OF THE INVENTION

Upon encountering antigen, naive CD4+ T helper precursor (Thp) cells aredifferentiated into two distinct subsets, Type 1 T helper (Th1) and Type2 T helper (Th2). Recently, a novel T cell subset, the Th17 cells, hasalso been identified and characterized These differentiated Th cells aredefined both by their distinct functional abilities and by uniquecytokine profiles. Specifically, Th1 cells produce interferon-gamma,interleukin (IL)-2, and tumor necrosis factor (TNF)-beta, which activatemacrophages and are responsible for cell-mediated immunity andphagocyte-dependent protective responses. In contrast, Th2 cells areknown to produce IL-4, IL-5, IL-6, IL-9, IL-10 and IL-13, which areresponsible for strong antibody production, eosinophil activation, andinhibition of several macrophage functions, thus providingphagocyte-independent protective responses. Th17 cells mainly produceIL-17A, IL-17F, IL-21, IL-22 & TNF and are required for host defenseagainst extracellular pathogens and are critical mediators ofautoimmunity. Accordingly, Th1, Th2, and Th17 cells are associated withdifferent immunopathological responses.

In addition, the development of each type of Th cell is mediated by adifferent cytokine pathway. Specifically, it has been shown that IL-4promotes Th2 differentiation and simultaneously blocks Th1 development.In contrast, IL-12, IL-18 and IFN-gamma are the cytokines critical forthe development of Th1 cells. In murine, TGF-β & IL-6 are critical forthe induction of Th17 cell differentiation, while in human, IL-1, IL-6 &IL-23 are important drivers of Th17 cell development. Accordingly,effective immunologic homeostasis relies on a continual balance betweenhelper T cell activation and regulatory T cell (Treg) suppression.

Th1 cells are involved in the pathogenesis of a variety oforgan-specific autoimmune disorders, Crohn's disease, Helicobacterpylori-induced peptic ulcer, acute kidney allograft rejection, andunexplained recurrent abortions. In contrast, allergen-specific Th2responses are responsible for atopic disorders in geneticallysusceptible individuals. Moreover, Th2 responses against still unknownantigens predominate in Omenn's syndrome, idiopathic pulmonary fibrosis,and progressive systemic sclerosis. Th17 cells cause immunopathology indifferent models of autoimmunity, such as rheumatoid arthritis, multiplesclerosis, Crohn's disease and psoriasis. IL-17 (the signature Th-17cytokine) knock-out mice show marked resistance to inflammatoryarthritis development. Joint destruction in the CIA model can beameliorated by the administration of a neutralizing anti-IL-17 antibody.

There remains a high unmet medical need to develop new therapeutictreatments that are useful in treating the various conditions associatedwith imbalanced Th1/Th2 and Th17 cellular differentiation. For many ofthese conditions the currently available treatment options areinadequate. Accordingly, the Th1/Th2 and Th17 paradigm provides arationale for the development of strategies for the therapy of allergicand autoimmune disorders.

Prostaglandins have been shown to modulate various phases of the immuneresponse. The lipid mediator prostaglandin E2 (PGE2) is an eicasanoidthat is well known to suppress CD4⁺ T cell activation through elevationof intracellular cAMP and inactivation of Ick. PGE2 has been also shownto play a role in regulating Th1 responses by suppression of interferongamma (IFN-gamma) production and T cell proliferation. However PGE2stimulation via the EP4 subtype of PGE2 receptor can also have theopposite effect, namely to promote Th1 differentiation (Prostaglandin Ereceptor subtypes EP2 and EP4 promote differentiation and expansion ofTh1 and Th17 lymphocytes through different signaling modules, NatureMedicine, 2009, 15, 633-640) and IL-17 production in activated CD4+cells. Prostaglandin E2 synergistically with interleukin-23 favors humanTh17 expansion, Blood, 2008, 112, 3696-3703; Prostaglandin E2 regulatesTh17 cell differentiation and function through cyclic AMP and EP2/EP4receptor signaling, J. Exp. Med. 2009, 206, 535-548; Prostaglandin E2enhances Th17 response via modulation of IL-17 and IFN-γ production bymemory CD4+ T cells, Eur. J. Immunol. 2009, 39, 1301-1312. Consistentwith this, antagonism of EP4 with either a novel selective EP4antagonist or a PGE2-neutralizing antibody suppresses Th1differentiation, Th17 expansion, as well as IL-23 secretion by activateddendritic cells. Induction of Th1 differentiation by PGE2 is mediated byPI3K signaling whereas stimulation of IL-17 production requires cAMPsignaling. In addition, administration of an EP4 antagonist to DBA/1 orC57BL/6 mice suppressed innate and adaptive immune responses, andsuppressed disease in collagen induced arthritis (CIA) and experimentalautoimmune encephalomyelitis (EAE) models, indicating that PGE2/EP4signaling is critically involved in these autoimmune pathologies. Theseresults suggest that suppression of PGE2/EP4 signaling may havetherapeutic value in modifying inflammatory autoimmune diseases such asrheumatoid arthritis and multiple sclerosis.

SUMMARY OF THE INVENTION

As described herein, the present invention provides compounds of FormulaI:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl (e.g., monofluoromethyl, difluoromethyl,trifluoromethyl);

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy (e.g.,monofluoromethoxy, difluoromethoxy, trifluoromethoxy);

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

In some embodiments, one of R_(1a) and R_(1b) is hydrogen, and the otheris methyl, and the carbon marked with a * is a stereogenic center. Insome embodiments, one of R_(1a) and R_(1b) is hydrogen, and the other ismethyl and the carbon marked with a * has the S-configuration. In someembodiments, one of R_(1a) and R_(1b) is hydrogen, and the other ismethyl and the carbon marked with a * has the R-configuration.

In some embodiments, the present invention provides a pharmaceuticalcomposition comprising a compound of Formula I or a subset or examplethereof. In some embodiments, the invention provides a method oftreating rheumatoid arthritis in a subject, comprising the step ofadministering to the subject a composition comprising a compound ofFormula I or a subset or example thereof. In some embodiments, theinvention provides a method of treating multiple sclerosis in a subject,comprising the step of administering to the subject a compositioncomprising a compound of Formula I or a subset or example thereof.

A further aspect of the invention is the use of a compound of Formula Ior a subset or example thereof in the manufacture of a medicament forthe treatment of rheumatoid arthritis. Another aspect of the inventionis the use of a compound of formula I or a subset or example thereof inthe manufacture of a medicament for the treatment of multiple sclerosis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Suppression of arthritis development in a CIA model with acompound of the present invention.

FIG. 2: Suppression of arthritis development in a glucose-6-phosphateisomerase (G6PI) model with a compound of the present invention.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION A.Definitions

Compounds of this invention include those described generally above, andare further illustrated by the embodiments, sub-embodiments, and speciesdisclosed herein. As used herein, the following definitions shall applyunless otherwise indicated.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. In general, the term “substituted” refersto the replacement of hydrogen radicals in a given structure with theradical of a specified substituent. Unless otherwise indicated, asubstituted group may have a substituent at each substitutable positionof the group, and when more than one position in any given structure maybe substituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds.

As used herein, the term “modulator of Th1 differentiation or Th17expansion” or “modulator compound of Th1 differentiation or Th17expansion” or “modulator compound” as used herein refers to a compoundwhich suppresses, reduces or inhibits, differentiation of naive CD4+ Tcells into Th1 cells. In some embodiments, the term “modulator of Th1differentiation or Th17 expansion” or “modulator compound of Th1differentiation or Th17 expansion” as used herein refers to a compoundwhich suppresses, reduces or inhibits, the number of IL-17 producingCD4+ T cells or IL-17 production in activated CD4+ T cells.

“Isomers” refer to compounds having the same number and kind of atomsand hence the same molecular weight, but differing with respect to thearrangement or configuration of the atoms.

“Stereoisomers” refer to isomers that differ only in the arrangement ofthe atoms in space.

“Diastereoisomers” refer to stereoisomers that are not mirror images ofeach other.

“Enantiomers” refers to stereoisomers that are non-superimposable mirrorimages of one another.

Enantiomers include “enantiomerically pure” isomers that comprisesubstantially a single enantiomer, for example, greater than or equal to90%, 92%, 95%, 98%, or 99%, or equal to 100% of a single enantiomer.

“Enantiomerically pure” as used herein means a compound, or compositionof a compound, that comprises substantially a single enantiomer, forexample, greater than or equal to 90%, 92%, 95%, 98%, or 99%, or equalto 100% of a single enantiomer.

“Stereomerically pure” as used herein means a compound or compositionthereof that comprises one stereoisomer of a compound and issubstantially free of other stereoisomers of that compound. For example,a stereomerically pure composition of a compound having one chiralcenter will be substantially free of the opposite enantiomer of thecompound. A stereomerically pure composition of a compound having twochiral centers will be substantially free of diastereomers, andsubstantially free of the enantiomer, of the compound. A typicalstereomerically pure compound comprises greater than about 80% by weightof one stereoisomer of the compound and less than about 20% by weight ofother stereoisomers of the compound, more preferably greater than about90% by weight of one stereoisomer of the compound and less than about10% by weight of the other stereoisomers of the compound, even morepreferably greater than about 95% by weight of one stereoisomer of thecompound and less than about 5% by weight of the other stereoisomers ofthe compound, and most preferably greater than about 97% by weight ofone stereoisomer of the compound and less than about 3% by weight of theother stereoisomers of the compound. See, e.g., U.S. Pat. No. 7,189,715.

“R” and “S” as terms describing isomers are descriptors of thestereochemical configuration at an asymmetrically substituted carbonatom. The designation of an asymmetrically substituted carbon atom as“R” or “S” is done by application of the Cahn-Ingold-Prelog priorityrules, as are well known to those skilled in the art, and described inthe International Union of Pure and Applied Chemistry (IUPAC) Rules forthe Nomenclature of Organic Chemistry. Section E, Stereochemistry.

“Enantiomeric excess” (ee) of an enantiomer is [(the mole fraction ofthe major enantiomer) minus (the mole fraction of the minorenantiomer)]×100.

“Stable”, as used herein, refers to compounds that are not substantiallyaltered when subjected to conditions to allow for their production,detection, and preferably their recovery, purification, and use for oneor more of the purposes disclosed herein. In some embodiments, a stablecompound or chemically feasible compound is one that is notsubstantially altered when kept at a temperature of 40° C. or less, inthe absence of moisture or other chemically reactive conditions, for atleast a week.

“Ar” or “aryl” refer to an aromatic carbocyclic moiety having one ormore closed rings. Examples include, without limitation, phenyl,naphthyl, anthracenyl, phenanthracenyl, biphenyl, and pyrenyl.

“Heteroaryl” refers to a cyclic moiety having one or more closed rings,with one or more heteroatoms (for example, oxygen, nitrogen or sulfur)in at least one of the rings, wherein at least one of the rings isaromatic, and wherein the ring or rings may independently be fused,and/or bridged. Examples include without limitation quinolinyl,isoquinolinyl, indolyl, furyl, thienyl, pyrazolyl, quinoxalinyl,pyrrolyl, indazolyl, thieno[2,3-c]pyrazolyl, benzofuryl,pyrazolo[1,5-a]pyridyl, thiophenylpyrazolyl, benzothienyl,benzothiazolyl, thiazolyl, 2-phenylthiazolyl, and isoxazolyl.

“Alkyl” or “alkyl group,” as used herein, means a straight-chain (i.e.,unbranched), branched, or cyclic hydrocarbon chain that is completelysaturated. In certain embodiments, alkyl groups contain 1-6 carbonatoms. In certain embodiments, alkyl groups contain 1-4 carbon atoms. Incertain embodiments, alkyl groups contain 1-3 carbon atoms. In stillother embodiments, alkyl groups contain 2-3 carbon atoms, and in yetother embodiments alkyl groups contain 1-2 carbon atoms. In certainembodiments, the term “alkyl” or “alkyl group” refers to a cycloalkylgroup, also known as carbocycle. Non-limiting examples of exemplaryalkyl groups include methyl, ethyl, propyl, isopropyl, butyl,cyclopropyl and cyclohexyl.

“Alkenyl” or “alkenyl group,” as used herein, refers to a straight-chain(i.e., unbranched), branched, or cyclic hydrocarbon chain that has oneor more double bonds. In certain embodiments, alkenyl groups contain 2-6carbon atoms In certain embodiments, alkenyl groups contain 2-4 carbonatoms. In still other embodiments, alkenyl groups contain 3-4 carbonatoms, and in yet other embodiments alkenyl groups contain 2-3 carbonatoms. According to another aspect, the term alkenyl refers to astraight chain hydrocarbon having two double bonds, also referred to as“diene.” In other embodiments, the term “alkenyl” or “alkenyl group”refers to a cycloalkenyl group. Non-limiting examples of exemplaryalkenyl groups include —CH═CH₂, —CH₂CH═CH₂ (also referred to as allyl),—CH═CHCH₃, —CH₂CH₂CH═CH₂, —CH₂CH═CHCH₃, —CH═CH₂CH₂CH₃, —CH═CH₂CH═CH₂,and cyclobutenyl.

“Alkoxy”, or “alkylthio”, as used herein, refers to an alkyl group, aspreviously defined, attached to the principal carbon chain through anoxygen (“alkoxy”) or sulfur (“alkylthio”) atom.

“Methylene”, “ethylene”, and “propylene” as used herein refer to thebivalent moieties —CH₂—, —CH₂CH₂—, and —CH₂CH₂CH₂—, respectively.

“Ethenylene”, “propenylene”, and “butenylene” as used herein refer tothe bivalent moieties —CH═CH—, —CH═CHCH₂—, —CH═CHCH₂CH₂—,—CH₂CH═CH₂CH₂—, and —CH₂CH₂CH═CH—, where each ethenylene, propenylene,and butenylene group can be in the cis or trans configuration. Incertain embodiments, an ethenylene, propenylene, or butenylene group canbe in the trans configuration.

“Alkylidene” refers to a bivalent hydrocarbon group formed by mono ordialkyl substitution of methylene. In certain embodiments, an alkylidenegroup has 1-6 carbon atoms. In other embodiments, an alkylidene grouphas 2-6, 1-5, 2-4, or 1-3 carbon atoms. Such groups include propylidene(CH₃CH₂CH═), ethylidene (CH₃CH═), and isopropylidene (CH₃(CH₃)CH═), andthe like.

“Alkenylidene” refers to a bivalent hydrocarbon group having one or moredouble bonds formed by mono or dialkenyl substitution of methylene. Incertain embodiments, an alkenylidene group has 2-6 carbon atoms. Inother embodiments, an alkenylidene group has 2-6, 2-5, 2-4, or 2-3carbon atoms. According to one aspect, an alkenylidene has two doublebonds. Exemplary alkenylidene groups include CH₂═CHCH═, CH₂═CHCH₂CH═,and CH₂═CHCH₂CH═CHCH═.

“C₁₋₆ alkyl ester or amide” refers to a C₁₋₆ alkyl ester or a C₁₋₆ alkylamide where each C₁₋₆ alkyl group is as defined above. Such C₁₋₆ alkylester groups are of the formula (C₁₋₆ alkyl)OC(═O)— or (C₁₋₆alkyl)C(═O)O—. Such C₁₋₆ alkyl amide groups are of the formula (C₁₋₆alkyl)NHC(═O)— or (C₁₋₆ alkyl)C(═O)NH—.

“C₂₋₆ alkenyl ester or amide” refers to a C₂₋₆ alkenyl ester or a C₂₋₆alkenyl amide where each C₂₋₆ alkenyl group is as defined above. SuchC₂₋₆ alkenyl ester groups are of the formula (C₂₋₆ alkenyl)OC(═O)— or(C₂₋₆ alkenyl)C(═O)O—. Such C₂₋₆ alkenyl amide groups are of the formula(C₂₋₆ alkenyl)NHC(═O)— or (C₂₋₆ alkenyl)C(═O)NH—.

““Fluoromethyl” as used herein refers to a methyl group substituted withone or more fluoro atoms (e.g., monofluoromethyl, difluoromethyl,trifluoromethyl).

“Fluoromethoxy” as used herein, refers to an fluoromethyl group, aspreviously defined, attached to the principal carbon chain through anoxygen atom.

“Treatment,” “treat,” and “treating” refer to reversing, alleviating,delaying the onset of, inhibiting the progress of, or preventing adisease or disorder as described herein. In some embodiments, treatmentmay be administered after one or more symptoms have developed. In otherembodiments, treatment may be administered in the absence of symptoms.For example, treatment may be administered to a susceptible individualprior to the onset of symptoms (e.g., in light of a history of symptomsand/or in light of genetic or other susceptibility factors). Treatmentmay also be continued after symptoms have resolved, for example toprevent or delay their recurrence.

“Patient” or “subject”, as used herein, means an animal subject,preferably a mammalian subject (e.g., dog, cat, horse, cow, sheep, goat,monkey, etc.), and particularly human subjects (including both male andfemale subjects, and including neonatal, infant, juvenile, adolescent,adult and geriatric subjects).

“Pharmaceutically acceptable carrier” as used herein refers to anontoxic carrier, adjuvant, or vehicle that does not destroy thepharmacological activity of the compound with which it is formulated.Pharmaceutically acceptable carriers, adjuvants or vehicles that may beused in the compositions of this invention include, but are not limitedto, ion exchangers, alumina, aluminum stearate, lecithin, serumproteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, cyclodextrins, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

“Pharmaceutically acceptable salt” refers to an acid or base salt of acompound of the invention, which salt possesses the desiredpharmacological activity and is neither biologically nor otherwiseundesirable. The salt can be formed with acids that include withoutlimitation acetate, adipate, alginate, aspartate, benzoate,benzenesulfonate, bisulfate butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, hydrochloride hydrobromide,hydroiodide, 2-hydroxyethane-sulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,thiocyanate, tosylate and undecanoate. Examples of a base salt includewithout limitation ammonium salts, alkali metal salts such as sodium andpotassium salts, alkaline earth metal salts such as calcium andmagnesium salts, salts with organic bases such as dicyclohexylaminesalts, N-methyl-D-glucamine, and salts with amino acids such as arginineand lysine. In some embodiments, the basic nitrogen-containing groupscan be quarternized with agents including lower alkyl halides such asmethyl, ethyl, propyl and butyl chlorides, bromides and iodides; diallylsulfates such as dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides; and aralkyl halides such as phenethyl bromides.

Unless indicated otherwise, nomenclature used to describe chemicalgroups or moieties as used herein follow the convention where, readingthe name from left to right, the point of attachment to the rest of themolecule is at the right-hand side of the name. For example, the group“(C₁₋₃ alkoxy)C₁₋₃ alkyl,” is attached to the rest of the molecule atthe alkyl end. Further examples include methoxyethyl, where the point ofattachment is at the ethyl end, and methylamino, where the point ofattachment is at the amine end.

Unless indicated otherwise, where a bivalent group is described by itschemical formula, including two terminal bond moieties indicated by “-,”it will be understood that the attachment is read from left to right.

Unless otherwise stated, structures depicted herein are also meant toinclude all enantiomeric, diastereomeric, and geometric (orconformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. Additionally, unless otherwise stated, structuresdepicted herein are also meant to include compounds that differ only inthe presence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools or probes inbiological assays.

B. Compounds

In one embodiment, the present invention provides a compound of FormulaI:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl (e.g., monofluoromethyl, difluoromethyl,trifluoromethyl);

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy (e.g.,monofluoromethoxy, difluoromethoxy, trifluoromethoxy);

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

In some embodiments, one of R_(1a) and R_(1b) is hydrogen, and the otheris methyl; R₂ is methyl, difluoromethyl, or trifluoromethyl; R₃ ismethyl; R₄ is chloro, fluoro, trifluoromethyl, difluoromethyl, methyl,methoxy, difluoromethoxy, or trifluoromethoxy; and R₅ is hydrogen,chloro, fluoro, methyl, or methoxy.

In some embodiments, R₅ is hydrogen.

In some embodiments, R₆ and R₇ are both hydrogen.

In some embodiments, one of R_(1a) and R_(1b) is hydrogen, and the otheris methyl, and R₄ is selected from chloro, trifluoromethyl,difluoromethyl, difluoromethoxy, and trifluoromethoxy.

In some embodiments, R_(1a) and R_(1b) are taken together to form acyclopropyl ring; R₂ is methyl, trifluoromethyl, or difluoromethyl; R₃is methyl; and R₄ is trifluoromethyl, difluoromethyl, chloro, or fluoro.

In some embodiments, one of R_(1a) and R_(1b) is hydrogen, and the otheris methyl, and the compound of Formula I consists of a mixture ofstereoisomers. In some embodiments, one of R_(1a) and R_(1b) ishydrogen, and the other is methyl, and the compound of Formula Iconsists of a substantially pure stereoisomer. In some embodiments, oneof R_(1a) and R_(1b) is hydrogen, and the other is methyl, and thecarbon of Formula I marked with a * has substantially theS-configuration. In some embodiments, one of R_(1a) and R_(1b) ishydrogen, and the other is methyl, and the carbon of Formula I markedwith a * has substantially the R-configuration.

C. Pharmaceutical Formulations

Active compounds of the present invention can be combined with apharmaceutically acceptable carrier to provide pharmaceuticalformulations thereof. The particular choice of carrier and formulationwill depend upon the particular route of administration for which thecomposition is intended.

The compositions of the present invention may be suitable for oral,parenteral, inhalation spray, topical, rectal, nasal, buccal, vaginal orimplanted reservoir administration, etc. Preferably, the compositionsare administered orally, intraperitoneally or intravenously. Sterileinjectable forms of the compositions of this invention may be aqueous oroleaginous suspension. These suspensions may be formulated according totechniques known in the art using suitable dispersing or wetting agentsand suspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a nontoxic parenterallyacceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

The pharmaceutically acceptable compositions of this invention may beorally administered in any orally acceptable dosage form including, butnot limited to, capsules, tablets, aqueous suspensions or solutions. Inthe case of tablets for oral use, carriers commonly used include lactoseand corn starch. Lubricating agents, such as magnesium stearate, arealso typically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

The pharmaceutically acceptable compositions of this invention may alsobe administered topically, especially when the target of treatmentincludes areas or organs readily accessible by topical application,including diseases of the eye, the skin, or the lower intestinal tract.Suitable topical formulations are readily prepared for each of theseareas or organs.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically transdermal patches may also be used.

For topical applications, the pharmaceutically acceptable compositionsmay be formulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutically acceptable compositions canbe formulated in a suitable lotion or cream containing the activecomponents suspended or dissolved in one or more pharmaceuticallyacceptable carriers. Suitable carriers include, but are not limited to,mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax,cetearyl alcohol, 2 octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutically acceptable compositions may beformulated as micronized suspensions in isotonic, pH adjusted sterilesaline, or, preferably, as solutions in isotonic, pH adjusted sterilesaline, either with or without a preservative such as benzylalkoniumchloride. Alternatively, for ophthalmic uses, the pharmaceuticallyacceptable compositions may be formulated in an ointment such aspetrolatum.

The pharmaceutically acceptable compositions of this invention may alsobe administered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

Most preferably, the pharmaceutically acceptable compositions of thisinvention are formulated for oral administration.

D. Subjects and Methods of Use

Prostaglandins have been shown to modulate various phases of the immuneresponse. The lipid mediator prostaglandin E2 (PGE2) is an eicasanoidthat is well known to suppress CD4⁺ T cell activation through elevationof intracellular cAMP and inactivation of Ick. PGE2 has been also shownto play a role in regulating Th1 responses by suppression of interferongamma (IFN-gamma) production and T cell proliferation. However PGE2stimulation via the EP4 subtype of PGE2 receptor can also have theopposite effect, namely to promote Th1 differentiation (Prostaglandin Ereceptor subtypes EP2 and EP4 promote differentiation and expansion ofTh1 and Th17 lymphocytes through different signaling modules, NatureMedicine, 2009, in press) and IL-17 production in activated CD4+ cells(Prostaglandin E2 synergistically with interleukin-23 favors human Th17expansion, Blood, 2008, 112, 3696-3703, Prostaglandin E2 regulates Th17cell differentiation and function through cyclic AMP and EP2/EP4receptor signaling, J. Exp. Med. 2009, 206, 535-548, Prostaglandin E2enhances Th17 response via modulation of IL-17 and IFN-γ production bymemory CD4+ T cells, Eur. J. Immunol. 2009, 39, 1301-1312). Consistentwith this, antagonism of EP4 with either a novel selective EP4antagonist or a PGE2-neutralizing antibody suppresses Th1differentiation, Th17 expansion, as well as IL-23 secretion by activateddendritic cells. Induction of Th1 differentiation by PGE2 is mediated byPI3K signaling whereas stimulation of IL-17 production requires cAMPsignaling. In addition, administration of an EP4 antagonist to DBA/1 orC57BL/6 mice suppressed innate and adaptive immune responses, andsuppressed disease in collagen induced arthritis (CIA) and experimentalautoimmune encephalomyelitis (EAE) models, indicating that PGE2/EP4signaling is critically involved in these autoimmune pathologies. Theseresults suggest that suppression of PGE2/EP4 signaling may havetherapeutic value in modifying inflammatory autoimmune diseases such asrheumatoid arthritis and multiple sclerosis.

Active compounds of the present invention may be administered topatients or subjects to treat a variety of different condition,particularly patients or subjects afflicted with:

(a) rheumatoid arthritis (see, e.g., Targeting rheumatoid arthritis andjoint inflammation in the mouse, J. Clin. Invest. 2002, 110, 651-658);Prostaglandin E2 exacerbates collagen-induced arthritis in mice throughthe inflammatory interleukin-23/interleukin-17 axis, Arthritis Rheum.2007. 56:2608-2619);

(b) multiple sclerosis (see, e.g., Narumiya, S. In The ProstanoidReceptors in Signaling Network of Chronic Inflammation—The Role of FP inBleomycin-induced Pulmonary Fibrosis and The Role of EP4 in ExperimentalAutoimmune Encephalomyelitis in Mice, Eicosanoids and ChronicInflammation, Montana, February, 2008), Keystone Symposia, Montana,February, 2008; Prostaglandin E receptor subtypes EP2 and EP4 promotedifferentiation and expansion of Th1 and Th17 lymphocytes throughdifferent signaling modules, Nature Medicine, 2009, in press);

(c) systemic lupus erythematosus (see, e.g., T-bet regulates IgG classswitching and pathogenic auto Ab production, Proc. Natl. Acad. Sci. USA2002, 99, 5545-50; Imbalance of Th1/Th2 transcription factors inpatients with lupus nephritis, Rheumatology (Oxford) 2006, 45, 951-7);

(d) type 1 diabetes (see, e.g., Identification of a novel type 1diabetes susceptibility gene, T-bet, Human Genetics 2004, 111, 177-84;T-bet controls autoaggressive CD8 lymphocyte response in type Idiabetes, J. Exp. Med. 2004, 199, 1153-62);

(e) psoriasis (see, e.g., A molecule solves psoriasis? Systemictherapies for psoriasis inducing interleukin 4 and Th2 responses, J.Mol. Med. 2003, 81, 471-80); The IL-23/Th17 axis in theimmunopathogenesis of psoriasis, J Invest Dermatol 2009,doi:10.1038/jid.2009.59;

(9 atherosclerosis (see, e.g., T-bet deficiency reduces athersclerosisand alters plaque antigen-specific immune responses, Proc. Natl. Acad.Sci. USA 2005, 102, 1596-601);

(g) Crohn's disease (see, e.g., IL-23/IL-17 immunity as a hallmark ofCrohn's disease, Inflamm Bowl Dis. 2009, 14, 1175-1184, Theproinflammatory effect of prostaglandin E2 in experimental inflammatorybowel disease is mediated through the IL-23-IL-17 axis, J. Immunol.2007, 178, 8138-8147);

h) inflammatory pain (see, e.g., Prostaglandin E2 receptor EP4contributes to inflammatory pain hypersensitivity, J. Pharmacol. Exp.Ther. 2006, 319, 1096-1103);

(i) neuropathic pain (see, e.g., Localisation and modulation ofprostanoid receptors EP1 and EP4 in the rat chronic constriction injurymodel of neuropathic pain, Eur. J. Pain 2007, 11, 605-613);

(j) migraine-associated pain (see, e.g., BGC20-1531, a novel, potent,and selective EP4 receptor antagonist: a putative new treatment formigraine headache, Br. J. Pharmacol. 2009, 156, 316-327).

(k) Spondyloarthropathies (see, e.g., Nonsteroidal AntiinflammatoryDrugs reduce radiographic progression in patients with ankylosingspondylitis, Arthritis Rhuem. 2005, 52, 1756-1765; Efficacy ofcelecoxib, a cyclooxygenase 2-specific inhibitor, in the treatment ofankylosing spondylitis: a six-week controlled study with comparisonagainst placebo and against a conventional nonsteroidal antiinflammatorydrug. Arthritis Rheum. 2001, 44, 180-185, Increased numbers ofcirculating polyfunctional Th17 memory cells in patients withseronegative spondylarthritides, Arthritis Rheum, 2008, 58, 2307-2317);

(l) Skin cancer (see, e.g., Chemoprevention of nonmelanoma skin cancerwith Celecoxib: A randomized, double-blind, placebo-controlled trial, JNatl Cancer Inst, 2010, 102, 1-10);

(m) Breast cancer (see, e.g., Potential new drug targets againsthormone-dependent breast cancer identified, Exp. Rev. Anticancer Ther.2008, 8, 507-509; Antagonism of the prostaglandin E receptor EP4inhibits metastasis and enhances NK function, Breast Cancer Res. Treat.2009, 117, 235-242; Prostaglandin E receptor EP4 antagonism inhibitsbreast cancer metastasis, Cancer Res. 2006, 66, 2923-2927);

(n) Colorectal cancer (see, e.g., Increased EP4 receptor expression incolorectal cancer progession promotes cell growth and anchorageindependence, Cancer Res. 2006, 66, 3106-3113);

(o) Prostate cancer (see, e.g., Identification of EP4 as a potentialtarget for the treatment of castration-resistant prostate cancer using anovel xenograft model, Cancer Res. 2010, 70, 1606-1615).

(p) Kidney cancer (see, e.g., Prostaglandin E2 regulates renal cellcarcinoma invasion through a EP4-Rap signal transduction pathway, J.Bio. Chem. Aug. 10, 2011 (epub)).

(q) Cervical cancer (see, e.g., COX-2 expression is correlated withVEGF-C, lymphangiogenesis and lymph node metastasis in human cervicalcancer, Microvasc Res. 2011, 82,131-40).

(r) Ovarian cancer (see, e.g., Ovarian epithelial cancer: a role forPGE2 synthesis and signaling in malignant transformation andprogression, Mol Cancer, 2006, 5, 62.)

(s) Endometrial cancer (see, e.g., Prostaglandin E2 inducesproliferation of Glandular epithelial cells of human endometrium viaextracellular regulated kinase 1/2-mediated patheway. J Clin Endocrinol& Metabol. 2003, 88, 4481-4487).

(t) Glioblastoma (see, e.g. Microsomal prostaglandin E synthase-1regulates human glioma cell growth via prostaglandin E₂-dependentactivation of type II protein kinase A. Mol. Cancer. Ther. 2006, 5,1817-1826).

(u) Head and neck cancer (see, e.g. Expression of prostaglandin E₂receptors in oral squamous cell carcinomas and growth inhibitory effectsof a selective EP3 antagonist, ONO-AE3-240. Int. J. Oncology 2009, 34,847-852).

(v) Medulloblastoma (see, e.g. Tumor-growth-promoting cyclooxygenase-2prostaglandin E2 pathway provides medulloblastoma therapeutic targets.Neuro-Oncol. 2008, 661-674).

(w) Lung cancer (see, e.g. Tumor cyclooxygenase-2/prostaglandinE2-dependent promotion of FOXP3 expression and CD4+ CD25+T regulatorycell activities in lung cancer. Cancer Res. 2005, 65, 5211-5220).

(x) Urinary tract cancers (see, e.g. Pathological function ofprostaglandin E₂ receptors in transitional cell carcinoma of the upperurinary tract. Virchows Archiv. 2006, 448, 822-829).

In addition, PGE2 has been implicated as an important consituent in theimmunosuppressive environment created by many solid tumors: Inhibitingthe inhibitors: evaluating agents targeting cancer immunosuppression.Expert Opinion in Biological Therapy. 2010. 10, 1019-35.EP4 receptorantagonism has been shown to reduce tumor metastasis: Host and directantitumor effects and profound reduction in tumor metastasis withselective EP4 receptor antagonism. Cancer Res. 2006, 66, 9665-9672.

Active compounds may be administered to subjects by any suitable route,including orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir.The term “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques. Preferably, the compositions are administeredorally, intraperitoneally or intravenously.

The active compounds are administered to the subjects in a treatmenteffective, or therapeutically effective, amount. The amount of thecompounds of the present invention that may be combined with the carriermaterials to produce a composition in a single dosage form will varydepending upon the host treated, and the particular route ofadministration. Preferably, the compositions should be formulated sothat a dosage of between 0, 01-100 mg/kg body weight/day of theinhibitor can be administered to a patient receiving these compositions.In certain embodiments, the compositions of the present inventionprovide a dosage of between 0.01 mg and 50 mg is provided. In otherembodiments, a dosage of between 0.1 and 25 mg or between 5 mg and 40 mgis provided.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of a compound of the present invention in the composition willalso depend upon the particular compound in the composition.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Examples 1-113 General

Microwave heating was done using Biotage Emrys Liberator or Initiatormicrowave. Column chromatography was carried out using Biotage SP4.Solvent removal was carried out using either a Büchii rotary evaporatoror a Genevac centrifugal evaporator. Preparative LC/MS was conductedusing a Waters autopurifier and 19×100 mm XTerra 5 micron MS C18 columnunder acidic mobile phase condition. NMR spectra were recorded usingVarian 400 MHz spectrometer.

When the term “inerted” is used to describe a reactor (e.g., a reactionvessel, flask, glass reactor, and the like) it is meant that the air inthe reactor has been replaced with an essentially moisture-free or dry,inert gas (such as nitrogen, argon, and the like).

General methods and experimentals for preparing compounds of the presentinvention are set forth below. In certain cases, a particular compoundis described by way of example. However, it will be appreciated that ineach case a series of compounds of the present invention were preparedin accordance with the schemes and experimentals described below.

The following abbreviations are used herein:

DEFINITIONS

The following abbreviations have the indicated meanings:

HATU: N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumHexafluorophosphate

COMU:(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate

DPCI: N,N′-Diisopropylcarbodiimide

DIEA: N,N-diisopropylethylamine

TEA: triethylamine

DMAP: 4-Dimethylaminopyridine

DMF: N,N-dimethylformamide

NMP: N-methylpyrrolidine

THF: tetrahydrofuran

DCM: dichloromethane

TFA: trifluoroacetic acid

Materials:

The following compounds are commercially available:

-   5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde (Maybridge Chemical    Co., Ltd.)-   5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde    (Maybridge Chemical Co., Ltd.)-   5-(3-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylic    acid (Bionet Research)-   1,3-dimethyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylic    acid (Bionet Research)-   1-methyl-3-(trifluoromethyl)-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4    carboxylic acid (Bionet Research)-   5-(4-chlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid    (Bionet Research)-   Ethyl 4,4-difluoroacetoacetate (Matrix Scientific)-   (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride (NetChem, Inc)-   4-(1-aminocyclopropyl)benzoic acid (Allweys LLC)    All phenols except for 3-difluoromethylphenol were commercially    available.    Compounds of the invention were made according to the general    synthetic scheme shown in Scheme I:

Preparation of representative non-limiting examples of the compounds ofthe invention are described below.

Examples 1-51 Production Scheme 1 Preparation procedure for5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde

Step 1:

Ethyl 4,4-difluoroacetoacetate (30.12 g, 0.172 mol) was stirred intoluene (600 mL) over ice water. Solution of N-methylhydrazine (7.6 mL,0.14 mol) in toluene (200 mL) was added slowly, dropwise over 20 min.The reaction mixture was heated at 100° C. for 2 hours. The reactionmixture evaporated to dryness. The resulting material was trituratedwith methyl t-butylether/heptane to give3-(difluoromethyl)-1-methyl-1H-pyrazol-5(4H)-one in three batches (total10.7 g, 51%) as orange powder. This material was used without furtherpurification for the next reaction.Step 2:

DMF (9.5 mL, 0.12 mol) was stirred over ice bath and phosphoryl chloride(24.0 mL, 0.257 mol) was added dropwise. To the solution was added3-(difluoromethyl)-1-methyl-1H-pyrazol-5(4H)-one (5.51 g, 0.0372 mol)portion wise and the mixture was heated at 120° C. for 40 minutes. Thereaction mixture was cooled, and the phosphoryl chloride was quenched byadding small chunks of ice slowly with stirring. The mixture was thenextracted with ethyl acetate three times and combined organic layer waswashed with water and brine, dried over MgSO4 and evaporated to give5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde (5.66 g,78.2%) as a dark orange/brown solid. This material was used withoutfurther purification for the next reaction.

Production Scheme 2 Preparation procedure for 3-difluoromethylphenol

To a solution of 3-hydroxybenzaldehyde (1.01 g, 0.0083 mol) in methylenechloride (3.3 mL) was added a solution of bis(2-methoxyethyl)aminosulfurtrifluoride (3.09 g, 0.0140 mol) in methylene chloride (3.3 mL) followedby ethanol (95.0 μL, 0.00163 mol). The reaction mixture was stirred atroom temperature for 12 hours. The mixture was then quenched carefullyadding saturated aqueous sodium bicarbonate solution. The organic layerseparated, washed with brine, dried over MgSO4, and evaporated to giveyellow oil. This oil was purified by column chromatography (0% to 50%ethyl acetate/heptane) to give the title compound (649 mg, 57%) ascolorless oil.

Production Scheme 3 Exemplary Procedure for the preparation of5-Aryloxy-pyrazole-4-carboxylic acid Production Example 15-(3,4-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

Step 1:

5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde (500.0 mg, 0.00315mol), 3,4-dichlorophenol (565 mg, 0.00344 mol) and potassium hydroxide(265 mg, 0.00472 mol) were stirred in DMF (2.0 mL). Mixture was heatedat 150° C. for 20 min using microwave. Water was added and resultingmixture was extracted with ethyl acetate. Organic layer was washed withbrine twice, dried over MgSO4 and evaporated. The resulting oil waspurified by column chromatography (10% to 20% ethyl acetate/heptane) togive 5-(3,4-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carbaldehyde(492 mg, 54.7%) as white solid.Step 2:

5-(3,4-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carbaldehyde fromStep 1 (492 mg, 0.001726 mol) and 2-methyl-2-butene (300 μL, 0.002832mol) were stirred in tert-butyl alcohol (2.0 mL). A solution of sodiumchlorite (400 mg, 0.003538 mol) and sodium dihydrogenphosphate (450 mg,0.003751 mol) in water (3.0 mL) was added and reaction mixture wasstirred for 12 hours at room temperature. Solvent was evaporated andresulting residue was dissolved in ethyl acetate. The organic layer waswashed with water, dried over MgSO4 and evaporated to give the titlecompound (340 mg, 65.5%) as white solid. This material was used in thenext step without further purification.

Production Example 25-(3-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3-fluorophenol inthe manner similar to the method in Production Example 1 above.

Production Example 35-(3,4-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3,5-difluorophenolin the manner similar to the method in Production Example 1 above.

Production Example 45-(2,3-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 2,3-difluorophenolin the manner similar to the method in Production Example 1 above.

Production Example 55-(2,5-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 2,5-difluorophenolin the manner similar to the method in Production Example 1 above.

Production Example 65-(3,5-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3,5-dichlorophenolin the manner similar to the method in Production Example 1 above.

Production Example 75-(3-chlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3-chlorophenol inthe manner similar to the method in Production Example 1 above.

Production Example 85-(2,3-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 2,3-dichlorophenolin the manner similar to the method in Production Example 1 above.

Production Example 95-(3-chloro-5-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and3-chloro-5-fluorophenol in the manner similar to the method inProduction Example 1 above.

Production Example 105-(2-chloro-5-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and2-chloro-5-fluorophenol in the manner similar to the method inProduction Example 1 above.

Production Example 111,3-dimethyl-5-(m-tolyloxy)-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and m-cresol in themanner similar to the method in Production Example 1 above.

Production Example 125-(3,5-dimethylphenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3,5-dimethylphenolin the manner similar to the method in Production Example 1 above.

Production Example 135-(3,5-dimethoxyphenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 3,5-dimethoxyphenolin the manner similar to the method in Production Example 1 above.

Production Example 145-(3-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and3-chlorophenol in the manner similar to the method in Production Example1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 153-(difluoromethyl)-1-methyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-4H-pyrazole-4-carbaldehyde and3-trifluoromethylphenol in the manner similar to the method inProduction Example 1 above except potassium carbonate was used insteadof potassium hydroxide.

Production Example 161,3-dimethyl-5-(3-(trifluoromethoxy)phenoxy)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and3-trifluoromethoxylphenol in the manner similar to the method inProduction Example 1 above.

Production Example 175-(3-fluorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and3-fluorophenol in the manner similar to the method in Production Example1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 185-(3,4-difluorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and3,4-difluorophenol in the manner similar to the method in ProductionExample 1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 195-(3,4-dichlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and3,4-dichlorophenol in the manner similar to the method in ProductionExample 1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 205-(3-(difluoromethoxy)phenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and3-difluoromethoxyphenol in the manner similar to the method inProduction Example 1 above.

Production Example 215-(3-(difluoromethoxy)phenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and3-difluoromethoxyphenol in the manner similar to the method inProduction Example 1 above except potassium carbonate was used insteadof potassium hydroxide.

Production Example 225-(3-(difluoromethoxy)phenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and3-difluoromethoxyphenol in the manner similar to the method inProduction Example 1 above except potassium carbonate was used insteadof potassium hydroxide.

Production Example 233-(difluoromethyl)-5-(3-(difluoromethyl)phenoxy)-1-methyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and3-difluoromethylphenol in the manner similar to the method in ProductionExample 1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 245-(3-(difluoromethyl)phenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and3-difluoromethylphenol in the manner similar to the method in ProductionExample 1 above.

Production Example 255-(3-(difluoromethyl)phenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and3-difluoromethylphenol in the manner similar to the method in ProductionExample 1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 265-(3,4-dichlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and3,4-dichlorophenol in the manner similar to the method in ProductionExample 1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 275-(4-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and 4-fluorophenol inthe manner similar to the method in Production Example 1 above.

Production Example 281,3-dimethyl-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and4-(trifluoromethyl)phenol in the manner similar to the method inProduction Example 1 above except cesium carbonate was used instead ofpotassium hydroxide.

Production Example 295-(2-chloro-4-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and2-chloro-4-fluorophenol in the manner similar to the method inProduction Example 1.

Production Example 301,3-dimethyl-5-(p-tolyloxy)-1H-pyrazole-4-carboxylic acid

The title compound was prepared using5-chloro-1,3-dimethyl-1H-pyrazole-4-carbaldehyde and p-cresol in themanner similar to the method in Production Example 1 above.

Production Example 311-methyl-3-(trifluoromethyl)-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and4-(trifluoromethyl)phenol in the manner similar to the method inProduction Example 1 above except potassium carbonate was used insteadof potassium hydroxide.

Production Example 323-(difluoromethyl)-1-methyl-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and4-(trifluoromethyl)phenol in the manner similar to the method inProduction Example 1 above except potassium carbonate was used insteadof potassium hydroxide.

Production Example 335-(4-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde and4-chlorophenol in the manner similar to the method in Production Example1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Example 345-(4-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid

The title compound was prepared using5-chloro-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carbaldehyde and4-chlorophenol in the manner similar to the method in Production Example1 above except potassium carbonate was used instead of potassiumhydroxide.

Production Scheme 4 Exemplary Procedure for Amide Coupling and EsterHydrolysis Example 1 ER-885289

5-(3-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid (50 mg, 0.0002 mol), HATU (65 mg, 0.00017 mol) and DIEA (30 μL,0.00017 mol) were stirred in DMF (1.0 mL) for 20 min at rt. Solution of(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride (40 mg, 0.00019 mol)and DIEA (32 μL, 0.00019 mol) in DMF (1.0 mL) was added dropwise and thereaction mixture was stirred at room temperature for 12 hours. Water wasadded and the resulting precipitate was collected, washed with water anddried under vacuum to give 59.2 mg of the methyl ester as white solid.This methyl ester was dissolved in methanol (1.0 mL) and THF (1.0 mL).2.0 M lithium hydroxide solution (240 μL) was added and the mixture wasstirred at room temperature for 12 hours. The reaction mixture wasneutralized with 1N hydrochloric acid solution (480 ul) and resultingemulsion was extracted with ethylacetate. The org. layer was separatedand evaporated. The resulting material was dissolved in 3 mL of methanoland was purified by LC/MS (0.1% TFA acetonitrile/water mobile phase).The desired fractions were evaporated by Genevac to give the titledcompound (27 mg, two steps 46%).

Example 2 ER-885290

Example 2 was prepared using1,3-dimethyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 1 above.

Example 3 ER-885291

Example 3 was prepared using1-methyl-3-(trifluoromethyl)-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 1 above.

Example 4 ER-885716

Example 4 was prepared using5-(3,4-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 1 above except hydrolysis was carried out in THF and1,4-dioxane at 140° C. for 10 min using microwave and product wasobtained as a solid after neutralization.

Example 5 ER-885717

Example 5 was prepared using5-(3-fluorophenoxy)-4,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 6 ER-885718

Example 6 was prepared using5-(3,4-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 7 ER-885719

Example 7 was prepared using5-(2,3-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 8 ER-885720

Example 8 was prepared using5-(2,5-difluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 9 ER-885721

Example 9 was prepared using5-(3,5-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 13 ER-885744

Example 13 was prepared using5-(3-chlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 14 ER-886022

Example 14 was prepared using5-(2,3-dichlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 15 ER-886024

Example 15 was prepared using5-(3-chloro-5-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acidand (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 16 ER-886025

Example 16 was prepared using5-(2-chloro-5-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acidand (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 17 ER-886032

Example 17 was prepared using1,3-dimethyl-5-(m-tolyloxy)-1H-pyrazole-4-carboxylic acid and (S)-methyl4-(1-aminoethyl)benzoate hydrochloride in a manner similar to theExample 4 above.

Example 18 ER-886033

Example 18 was prepared using5-(3,5-dimethylphenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 19 ER-886035

Example 19 was prepared using5-(3,5-dimethoxyphenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 20 ER-886045

Example 20 was prepared using5-(3-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 21 ER-886046

Example 21 was prepared using3-(difluoromethyl)-1-methyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 22 ER-886061

Example 22 was prepared using1,3-dimethyl-5-(3-(trifluoromethoxy)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 23 ER-886072

Example 23 was prepared using5-(3-fluorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 1 above except COMU, TEA and NMP was used insteadof HATU, DIEA and DMF, and the product was obtained as solid afterneutralization.

Example 24 ER-886073

Example 24 was prepared using5-(3,4-difluorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 23 above.

Example 25 ER-886074

Example 25 was prepared using5-(3,4-dichlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 23 above.

Example 26 ER-886077

Example 26 was prepared using5-(3-chlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid andmethyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a manner similarto the Example 4 above.

Example 27 ER-886078

Example 27 was prepared using3-(difluoromethyl)-1-methyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 28 ER-886080

Example 28 was prepared using5-(3-(difluoromethoxy)phenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 23 above.

Example 29 ER-886082

Example 29 was prepared using5-(3-(difluoromethoxy)phenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 23 above.

Example 30 ER-886083

Example 30 was prepared using3-(difluoromethyl)-1-methyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (R)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 31 ER-886090

Example 31 was prepared using5-(3-(difluoromethoxy)phenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 23 above.

Example 32 ER-887480

Example 32 was prepared using3-(difluoromethyl)-5-(3-(difluoromethyl)phenoxy)-1-methyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above except product was obtained after LC/MSpurification.

Example 33 ER-887495

Example 33 was prepared using5-(3-(difluoromethyl)phenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acidand (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above except HBTU, TEA and NMP was used insteadof HATU, DIEA and DMF.

Example 34 ER-887995

Example 34 was prepared using5-(3-(difluoromethyl)phenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 33 above except product was obtained after LC/MSpurification.

Example 35 ER-888024

Example 35 was prepared using5-(3,4-dichlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 36 ER-888348

Example 36 was prepared using5-(3,4-dichlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 37 ER-888355

Example 37 was prepared using5-(3-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 38 ER-888363

5-(3,4-dichlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid (104 mg, 0.0002929 mol), DMAP (17 mg, 0.000139 mol), PS-HOBT (1.00mmol/g loading; 191 mg, 0.000191 mol), were stirred in 20% DMF in DCM(4:1, DCM:DMF, 5.00 mL). DPCI (140 uL, 0.0008941 mol) was added and themixture was shaken at 40° C. 12 hours. The resin was then washedsequentially with DCM (3 ml), DMF (3 mL), DCM (3 ml), THF (3 ml), andthen again with DCM (3 ml), and dried under vacuume. This resin wasstirred in 20% DMF in DCM (4:1, DCM:DMF, 5.00 mL) and solution of methyl4-(1-aminocyclopropyl)benzoate hydrochloride (27 mg, 0.000118 mol) in20% DMF in DCM (5.00 mL) and DIEA (20 uL, 0.000115 mol) were added. Themixture was shaken at 40° C. for 12 hours. The resin was filtered andwas washed with DCM. The combined filtrates were concentrated in vacuoto give a DMF solution. The solution was diluted with ethyl acetatewashed with water and brine, dried over MgSO4 and evaporated to give abeige solid. The solid was purified by column chromatography (0 to 40%)to give methyl ester (18 mg) as a white solid. This methyl ester wasdissolved in 1,4-dioxane (1.0 mL). 2.0 M lithium hydroxide solution (200μL) was added and the mixture was heated at 60° C. for 12 hours. Thereaction mixture was further heated at 140° C. for 20 min usingmicrowave. The reaction mixture was acidified with 1N hydrochloric acidsolution (420 ul) and water was added. The resulting precipitate wascollected, washed with water and air dried to give the title compound(15.9 mg, two steps 26%) as white solid.

Example 39 ER-880663

Example 39 was prepared using5-(4-chlorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 40 ER-885302

Example 40 was prepared using5-(4-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acid and(S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a manner similar tothe Example 4 above.

Example 41 ER-885311

Example 41 was prepared using1,3-dimethyl-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 42 ER-886023

Example 42 was prepared using5-(2-chloro-4-fluorophenoxy)-1,3-dimethyl-1H-pyrazole-4-carboxylic acidand (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 4 above.

Example 43 ER-885749

Example 43 was prepared using1,3-dimethyl-5-(p-tolyloxy)-1H-pyrazole-4-carboxylic acid and (S)-methyl4-(1-aminoethyl)benzoate hydrochloride in a manner similar to theExample 4 above.

Example 44 ER-888365

Example 44 was prepared using1-methyl-3-(trifluoromethyl)-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 38.

Example 45 ER-888367

Example 45 was prepared using3-(difluoromethyl)-1-methyl-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 38 above except hydrolysis was carried out at 60°C. for 12 hours.

Example 46 ER-888369

Example 46 was prepared using5-(4-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 38 above except the product was obtained afterpurification using heptane/ethyl acetate/formic acid as mobile phase.

Example 47 ER-888371

Example 47 was prepared using5-(4-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and (S)-methyl 4-(1-aminoethyl)benzoate hydrochloride in a mannersimilar to the Example 38 above.

Example 48 ER-888364

Example 48 was prepared using1-methyl-3-(trifluoromethyl)-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 38 above.

Example 49 ER-888366

Example 49 was prepared using3-(difluoromethyl)-1-methyl-5-(4-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 38 above.

Example 50 ER-888368

Example 50 was prepared using5-(4-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 38 above.

Example 51 ER-888370

Example 51 was prepared using5-(4-chlorophenoxy)-3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylicacid and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride in a mannersimilar to the Example 38 above.

Example 10 ER-885740

1,3-dimethyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid (20 mg, 0.00007 mol), HATU (28 mg, 0.000073 mol) and DIEA (13 μL,0.000073 mol) were stirred in DMF (0.5 mL) for 20 min at rt. Solution of4-(1-aminocyclopropyl)benzoic acid (14 mg, 0.000080 mol) and DIEA (14μL, 0.000080 mol) in DMF (0.5 mL) was added dropwise and the reactionmixture was stirred at room temperature for 12 hours. The reactionmixture was diluted to 3 mL with methanol and purified by LC/MS (0.1%TFA Acetonitrile/water mobile phase). The desired fractions wereevaporated by Genevac to give the title compound (15 mg, 50%).

Example 11 ER-885741

Example 11 was prepared using1-methyl-3-(trifluoromethyl)-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4-carboxylicacid and 4-(1-aminocyclopropyl)benzoic acid in a manner similar to theExample 10 above.

Example 12 ER-885743

Example 12 was prepared using5-(3-chlorophenoxy)-1-methyl-3-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid and 1 4-(1-aminocyclopropyl)benzoic acid in a manner similar to theExample 10 above.

Preparation procedure for methyl 4-(1-aminocyclopropyl)benzoatehydrochloride

Step 1:

4-(1-aminocyclopropyl)benzoic acid (1.75 g, 0.00988 mol) was stirred inTHF (20 mL) and Methanol (10 mL) over ice/water bath. 2.00 M oftrimethylsilyldiazomethane in hexane (9.9 mL, 0.020 mol) was added andthe mixture was stirred at room temperature for 12 hours. The reactionmixture was concentrated down to viscous oil which solidified uponstanding to give a crystalline solid. Crude material was dried on highvacuum line for 12 hours to give methyl 4-(1-aminocyclopropyl)benzoate(1.65 g, 87%) as pale brown solid.Step 2:

To a solution of ER-886774-00 (1.63 g, 0.0085 mol) in ethyl acetate (10mL) was added 2.0 M of hydrogen chloride in Ether (6.0 mL, 0.012 mol).After stirring for several minutes, the reaction mixture wasconcentrated to give the title compound (quantitative yield) as palebrown solid.

TABLE 1 Analytical Data for Exemplary Compounds of Formula I ExampleStructure ER number 1H NMR assignment 1

ER-885289-00 1H NMR (400 MHz, CD3OD) δ ppm 8.56 (1H, d), 7.87 (2H, m),7.30 (1H, m), 7.18 (2H, d), 7.08 (1H, t), 6.88 (1H, m), 4.96 (1H, m),3.78 (3H, s), 1.34 (3H, d) 2

ER-885290-00 1H NMR (400 MHz, CD3OD) δ ppm 7.88 (1H, d), 7.80 (2H, d),7.49 (2H, m), 7.30 (1H, br. s.), 7.13 (2H, d), 4.99 (1H, m), 3.64 (3H,s), 2.37 (3H, m), 1.32 (3H, d) 3

ER-885291-00 1H NMR (400 MHz, CD3OD) δ ppm 8.60 (1H, d), 7.81 (2H, d),7.51 (2H, m), 7.39 (1H, s), 7.18 (3H, m), 4.92 (1H, m), 3.79 (3H, s),1.30 (3H, d) 4

ER-885716-00 1H NMR (400 MHz, CD3OD) δ ppm 7.84 (2H, d), 7.42 (1H, d),7.15 (2H, d), 7.13 (1H, d), 6.83 (1H, dd), 5.01 (1H, q), 3.62 (3H, s),2.35 (3H, s), 1.36 (3H, d) 5

ER-885717-00 1H NMR (400 MHz, CD3OD) δ ppm 7.8 (3H, m), 7.34 (1H, m),7.14 (2H, d), 6.91 (1H, m), 6.73 (2H, m), 5 (1H, q), 3.61 (3H, s), 2.35(3H, s), 1.34 (3H, d) 6

ER-885718-00 1H NMR (400 MHz, CD3OD) δ ppm 7.86 (3H, m), 7.19 (3H, m),6.92 (1H, m), 6.68 (1H, m), 5.02 (1H, q), 3.61 (3H, s), 2.34 (3H, s),1.37 (3H, d) 7

ER-885719-00 1H NMR (400 MHz, CD3OD) δ ppm 8.01 (1H, d), 7.84 (2H, d),7.19 (2H, d), 7.01 (2H, m), 6.58 (1H, m), 4.99 (1H, q), 3.66 (3H, s),2.33 (3H, s), 1.37 (3H, d) 8

ER-885720-00 1H NMR (400 MHz, CD3OD) δ ppm 8.06 (1H, d), 7.85 (2H, d),7.22 (3H, m), 6.87 (1H, m), 6.63 (1H, m), 5 (1H, q), 3.66 (3H, s), 2.32(3H, s), 1.37 (3H, d) 9

ER-885721-00 1H NMR (400 MHz, CD3OD) δ ppm 7.97 (1H, d), 7.85 (2H, d),7.18 (3H, m), 6.89 (1H, d), 5.03 (1H, q), 3.62 (3H, s), 2.35 (3H, s),1.38 (3H, d) 10

ER-885740-00 1H NMR (400 MHz, CD3OD) δ ppm 8.45 (1H, s), 7.79 (2H, d),7.59 (1H, m), 7.52 (1H, m), 7.36 (1H, s), 7.21 (1H, dd), 7.03 (2H, d),3.67 (3H, s), 2.38 (3H, s), 1.22 (2H, m), 0.91 (2H, m) 11

ER-885741-00 1H NMR (400 MHz, CD3OD) δ ppm 9.03 (1H, s), 7.77 (2H, d),7.58 (2H, m), 7.46 (1H, s), 7.26 (1H, d), 7.03 (2H, d), 3.83 (3H, s),1.20 (2H, m), 0.84 (2H, m) 12

ER-885743-00 1H NMR (400 MHz, CD3OD) δ ppm 7.80 (2H, d), 7.39 (1H, m),7.26 (1H, d), 7.17 (1H, t), 7.04 (2H, d), 6.97 (1H, dd), 3.82 (3H, s),1.23 (2H, m), 0.92 (2H, m) 13

ER-885744-00 1H NMR (400 MHz, CD3OD) δ ppm 7.83 (2H, d), 7.32 (1H, m),7.19 (1H, m), 7.14 (2H, d), 7.00 (1H, t), 6.85 (1H, dd), 5.01 (1H, m),3.63 (3H, s), 2.36 (3H, s), 1.35 (3H, d) 14

ER-886022-00 1H NMR (400 MHz, CD3OD) δ ppm 7.81 (2H, d), 7.72 (1H, d),7.31 (1H, d), 7.14 (3H, m), 6.64 (1H, d), 5 (1H, q), 3.63 (3H, s), 2.35(3H, s), 1.37 (3H, d) 15

ER-886024-00 1H NMR (400 MHz, CD3OD) δ ppm 7.96 (1H, d), 7.85 (2H, d),7.19 (2H, d), 6.98 (1H, dt), 6.79 (1H, m), 6.67 (1H, dt), 5.03 (1H, q),3.62 (3H, s), 2.35 (3H, s), 1.38 (3H, d) 16

ER-886025-00 1H NMR (400 MHz, CD3OD) δ ppm 7.83 (3H, m), 7.49 (1H, dd),7.18 (2H, m), 6.91 (1H, m), 6.53 (1H, dd), 5.01 (1H, q), 3.64 (3H, s),2.34 (3H, s), 1.38 (3H, d) 17

ER-886032-00 1H NMR (400 MHz, CD3OD) δ ppm 7.78 (2H,d), 7.53 (1H, d),7.24 (1H, t), 7.07 (1H, m), 7.00 (1H, d), 6.72 (2H, m), 4.99 (1H, m),3.59 (3H, s), 2.35 (3H, s), 2.29 (3H, s), 1.32 (3H, d) 18

ER-886033-00 1H NMR (400 MHz, CD3OD) δ ppm 7.77 (2H, d), 7.47 (1H, d),7.05 (2H, d), 6.83 (1H, s), 6.52 (2H, s), 5.00 (1H, m), 3.58 (3Hs), 2.35(3H, s), 2.24 (3H, s), 1.33 (3H, d) 19

ER-886035-00 1H NMR (400 MHz, CD3OD) δ ppm 7.79 (2H, d), 7.11 (2H, d),6.29 (1H, t), 6.04 (2H, d), 5.00 (1H, m), 3.71 (6H, s), 3.60 (3H, s),2.35 (3H, s), 1.36 (3H, d) 20

ER-886045-00 1H NMR (400 MHz, CD3OD) δ ppm 8.05 (1H, m), 7.84 (2H, d),7.32 (1H, t), 7.20 (1H, m), 7.15 (2H, d), 6.96 (3H, m), 4.99 (1H, q),3.75 (3H, s), 1.36 (3H, d) 21

ER-886046-00 1H NMR (400 MHz, CD3OD) δ ppm 8.11 (1H, m), 7.81 (2H, d),7.52 (2H, m), 7.37 (1H, s), 7.15 (3H, m), 6.96 (1H, m), 4.97 (1H, q),3.76 (3H, s), 1.32 (3H, d) 22

ER-886061-00 1H NMR (400 MHz, CD3OD) δ ppm 7.83 (3H, m), 7.43 (1H, t),7.12 (3H, m), 6.95 (1H, s), 6.87 (1H, dd), 5.00 (1H, quin), 3.63 (3H,s), 2.36 (3H, s), 1.33 (3H, d) 23

ER-886072-00 1H NMR (400 MHz, CD3OD) δ ppm 8.56 (1H, d), 7.82 (2H, d),7.33 (1H, m), 7.18 (2H, d), 6.91 (1H, m), 6.79 (2H, m), 4.95 (1H, m),3.77 (3H, s), 1.34 (3H, d) 24

ER-886073-00 1H NMR (400 MHz, CD3OD) δ ppm 8.60 (1H, d), 7.86 (2H, d),7.19 (3H, m), 6.99 (1H, m), 6.75 (1H, m), 4.97 (1H, m), 3.77 (3H, s),1.35 (3H, d) 25

ER-886074-00 1H NMR (400 MHz, CD3OD) δ ppm 8.61 (1H, d), 7.86 (2H, d),7.40 (1H, d), 7.22 (1H, d), 7.18 (2H, d), 6.91 (1H, m), 6.89 (1H, dd),4.96 (1H, m), 3.78 (3H, s), 1.35 (3H, d) 26

ER-886077-00 1H NMR (400 MHz, CD3OD) δ ppm 8.40 (1H, s), 7.81 (2H, d),7.38 (1H, t), 7.24 (1H, m), 7.06 (3H, m), 6.92 (1H, dd), 3.66 (3H, s),2.37 (3H, s), 1.24 (2H, m), 0.98 (2H, m) 27

ER-886078-00 1H NMR (400 MHz, CD3OD) δ ppm 7.78 (2H, d), 7.59 (2H, m),7.44 (1H, s), 7.24 (1H, m), 7.00 (3H, m), 3.80 (3H, s), 1.21 (2H, m),0.89 (2H, m) 28

ER-886080-00 1H NMR (400 MHz, CD3OD) δ ppm 7.81 (2H, d), 7.77 (1H, d),7.35 (1H, t), 7.14 (2H, d), 6.95 (1H, m), 6.80 (1H, t), 6.78 (1H, m),6.73 (1H, m), 5.00 (1H, m), 3.61 (3H, s), 2.35 (3H, s), 1.33 (3H, s) 29

ER-886082-00 1H NMR (400 MHz, CD3OD) δ ppm 8.55 (1H, d), 7.82 (2H, d),7.34 (1H, t), 7.17 (2H, d), 6.95 (1H, m), 6.83 (1H, t), 6.80 (1H, t),6.78 (2H, dd), 4.94 (1H, m), 3.76 (3H, s), 3.65 (3H, s), 1.32 (3H, s) 30

ER-886083-00 1H NMR (400 MHz, CD3OD) δ ppm 8.10 (1H, d), 7.81 (2H, d),7.52 (2H, m), 7.37 (1H, m), 7.08 (4H, m), 4.97 (1H, q), 3.76 (3H, s),1.32 (3H, d) 31

ER-886090-00 1H NMR (400 MHz, CD3OD) δ ppm 8.02 (1H, d), 7.81 (2H, d),7.36 (1H, t), 7.15 (2H, d), 6.98 (1H, m), 6.83 (1H, m), 6.81 (1H, t),6.76 (2H, dd), 4.99 (1H, m), 3.73 (3H, s), 1.34 (3H, s) 32

ER-887480-00 1H NMR (400 MHz, CD3OD) δ ppm 8.01 (1H, d), 7.79 (2H, d),7.46 (1H, t), 7.36 (1H, d), 7.19 (1H, s), 7.11 (2H, d), 7.06 (1H, m),6.96 (1H, t), 6.73 (1H, t), 4.95 (1H, m), 3.74 (3H, s), 1.31 (3H, s) 33

ER-887495-00 1H NMR (400 MHz, CD3OD) δ ppm 7.77 (3H, m), 7.46 (1H, t),7.34 (1H, d), 7.14 (1H, s), 7.10 (1H, d), 7.04 (1H, m), 6.73 (1H, t),4.97 (1H, q), 3.62 (3H, s), 2.35 (3H, s), 1.31 (3H, d) 34

ER-887995-00 1H NMR (400 MHz, CD3OD) δ ppm 8.53 (1H, d), 7.79 (2H, d),7.43 (1H, t), 7.34 (1H, d), 7.20 (1H, s), 7.13 (2H, d), 7.06 (1H, m),6.72 (1H, t), 4.91 (1H, m), 3.76 (3H, s), 1.28 (3H, d) 35

ER-888024-00 1H NMR (400 MHz, CD3OD) δ ppm 8.12 (1H, d), 7.86 (2 H, d),7.44 (1 H, d), 7.17 (5 H, d), 5.00 (1 H, q), 3.76 (3 H, s), 1.38 (3 H,d) 36

ER-888348-00 1H NMR (400 MHz, CD3OD) δ ppm 8.65 (1H, s), 7.81 (2H, d),7.52 (1H, d), 7.30 (1H, d), 7.03 (2H, d), 6.97 (1H, m), 6.91 (1H, t),3.77 (3H, s), 1.24 (2H, dd), 0.99 (2H, dd) 37

ER-888355-00 1H NMR (400 MHz, CD3OD) δ ppm 8.63 (1H, s), 7.79 (2H, d),7.38 (1H, t), 7.25 (1H, m), 7.14 (1H, t), 7.03 (2H, d), 6.95 (1H, m),6.93 (1H, t), 3.77 (3H, s), 1.23 (2H, dd), 0.96 (2H, dd) 38

ER-888363-00 1H NMR (400 MHz, CD3OD) δ ppm 7.81 (2H, d), 7.52 (1H, d),7.33 (1H, d), 7.06 (2H, d), 7.00 (1H, dd), 3.82 (3H, s), 1.24 (2H, m),0.95 (2H, m) 39

ER-880663-00 1H NMR (400 MHz, CD3OD) δ ppm 7.83 (2 H, s), 7.74 (1 H, d),7.33 (2 H, m), 7.13 (2 H, d), 6.93 (2 H, m), 5.01 (1 H, m), 3.62 (3 H,s), 2.35 (3 H, s), 1.35 (3H, d 40

ER-885302-00 1H NMR (400 MHz, CD3OD) δ ppm 7.78 (3 H, m) 7.05 (6 H, m)5.01 (1 H, m) 3.62 (3 H, s) 2.35 (3 H, s) 1.35 (3 H, d) 41

ER-885311-00 1H NMR (400 MHz, CD3OD) δ ppm 7.75 (5 H, m) 7.12 (4 H, m)5.01 (1 H, m) 3.63 (3 H, s) 2.37 (3 H, s) 1.33 (3 H, d) 42

ER-886023-00 1H NMR (400 MHz, CD3OD) δ ppm 7.83 (2H, m), 7.30 (1H, m),7.17 (2H, m), 6.95 (1H, m), 6.73 (1H, m), 5.00 (1H, m), 3.63 (3H, s),2.33 (3H, s), 1.38 (3H, d) 43

ER-885749-00 1H NMR (400 MHz, CD3OD) δ ppm 7.77 (2H, d), 7.54 (1H, d),7.14 (2H, d), 7.06 (2H, d), 6.80 (2H, d), 4.99 (1H, p), 3.59 (3H, s),2.34 (3H, s), 2.32 (3H, s), 1.32 (3H, d) 44

ER-888365-00 1H NMR (400 MHz, CD3OD) δ ppm 8.60 (1H, d), 7.81 (2H, d),7.64 (2H, d), 7.15 (4H, m), 4.94 (1H, m), 3.77 (3H, s), 1.31 (3H, d) 45

ER-888367-00 1H NMR (400 MHz, CD3OD) δ ppm 8.07 (1H, d), 7.81 (2H, d),7.66 (2H, d), 7.14 (4H, m), 6.96 (1H, t), 4.99 (1H, m), 3.75 (3H, s),1.34 (3H, d) 46

ER-888369-00 1H NMR (400 MHz, CD3OD) δ ppm 8.53 (1H, d), 7.85 (2H, d),7.30 (2H, d), 7.17 (2H, d), 6.97 (2H, d), 4.95 (1H, m), 3.77 (3H, s),1.34 (3H, d) 47

ER-888371-00 1H NMR (400 MHz, CD3OD) δ ppm 7.99 (1H, d), 7.85 (2H, d),7.33 (2H, d), 7.15 (2H, d), 6.96 (3H, m), 4.99 (1H, m), 3.74 (3H, s),1.36 (3H, d) 48

ER-888364-00 1H NMR (400 MHz, CD3OD) δ ppm 9.02 (1H, s), 7.76 (4H, m),7.24 (2H, d), 7.03 (2H, d), 3.82 (3H, s), 1.21 (2H, m), 0.88 (2H, m) 49

ER-888366-00 1H NMR (400 MHz, CD3OD) δ ppm 8.64 (1H, s), 7.76 (4H, m),7.22 (2H, d), 7.04 (2H, d), 6.95 (1H, t), 3.78 (3H, s), 1.23 (2H, m),0.94 (2H, m) 50

ER-888368-00 1H NMR (400 MHz, CD3OD) δ ppm 7.81 (2H, d), 7.40 (2H, d),7.04 (4H, m), 3.81 (3H, s), 1.23 (2H, m), 0.94 (2H, m) 51

ER-888370-00 1H NMR (400 MHz, CD3OD) δ ppm 7.81 (2H, d), 7.41 (2H, d),7.03 (4H, m), 6.94 (1H, t), 3.77 (3H, s), 1.24 (2H, m), 1.00 (2H, m)

Examples 52-111 In Vitro Biological Activity

CRE-PLAP Reporter Assay:

SE302 is a clone of the HEK/293 cell line that harbors a reporterconstruct containing a promoter with cAMP response elements (CRE)driving secreted alkaline phosphatase (PLAP), constructed by T. Arai,Eisai Pharamceuticals, Japan. These cells express endogenous EP4 andshow induction of PLAP in response to PGE2 and other agonists of EP4,but not of EP1, 2 or 3 (data not shown). Cells were maintained inDMEM/F12 (50:50) (MediaTech) supplemented with 10% FBS (Tissue CultureBiologicals) plus penicillin/streptomycin. When used for assays, cellswere plated in a 96-well plate at 2×10⁴ cells/100 μL/well in serum-freeassay medium (DMEM/F12 supplemented with 0.1% BSA pluspenicillin/streptomycin) and incubated for 4-6 h. Cells were thenstimulated with 3 ng.mL⁻¹ of PGE2 in the presence or absence of variousconcentrations of ER-819762 overnight, and PLAP activity was measured bymixing 15 μL of culture supernatants with 75 μL of Lumi-phos (Lumigen,Inc.) and 60 μL of assay buffer containing 8 mmol.L⁻¹ MgSO₄ in 0.1mol.L⁻¹ carbonate-bicarbonate buffer pH11 in a new 96-well black plateand incubated for 2 h at room temperature. Luminescence was read with anEnvision 2102 Multilabel reader.

Exemplary compounds of the present invention were assayed according tothe methods set forth above in the CRE-PLAP reporter assay describedabove. Table 2 below sets forth exemplary compounds of the presentinvention having an IC₅₀ of up to 5.0 μM as determined by the normalizedCRE-PLAP assay described above.

TABLE 2 IC₅₀ Values Exemplary Compounds CRE-PLAP Example Structure ERnumber IC50 (μM) 52

ER-885289-00 0.045 53

ER-885290-00 0.018 54

ER-885291-00 0.027 55

ER-885716-00 0.092 56

ER-885717-00 0.974 57

ER-885718-00 0.467 58

ER-885719-00 2.666 59

ER-885720-00 2.219 60

ER-885721-00 0.846 61

ER-885740-00 0.012 62

ER-885741-00 0.013 63

ER-885743-00 0.038 64

ER-885744-00 0.168 65

ER-886022-00 0.719 66

ER-886024-00 0.637 67

ER-886025-00 4.914 68

ER-886032-00 0.356 69

ER-886033-00 2.122 70

ER-886035-00 3.665 71

ER-886045-00 0.018 72

ER-886046-00 0.011 73

ER-886061-00 0.089 74

ER-886072-00 0.132 75

ER-886073-00 0.109 76

ER-886074-00 0.01 77

ER-886077-00 0.085 78

ER-886078-00 0.009 79

ER-886080-00 0.058 80

ER-886082-00 0.088 81

ER-886083-00 0.827 82

ER-886090-00 0.017 83

ER-887480-00 0.051 84

ER-887495-00 0.219 85

ER-887995-00 0.187 86

ER-888024-00 0.031 87

ER-888348-00 0.017 88

ER-888355-00 0.157 89

ER-888363-00 0.008 90

ER-880663-00 0619 91

ER-885302-00 0.261 92

ER-885311-00 0.273 93

ER-886023-00 4.403 94

ER-885749-00 0.451 95

ER-888365-00 0.028 96

ER-888367-00 0.023 97

ER-888369-00 0.089 98

ER-888371-00 0.089 99

ER-888364-00 0.012 100

ER-888366-00 0.015 101

ER-888368-00 0.018 102

ER-888370-00 0.035

Radioligand EP4 Receptor Binding Assay:

The radioligand EP₄ binding assay was performed using ChemiScreenrecombinant human EP₄ receptor membrane preparations from Millipore,according to manufacturer's instructions. Briefly, membranes preparedfrom Chem-1 cells overexpressing human EP₄ cDNA (Millipore) were mixedwith 1.8 nmol.L⁻¹ [³H]-PGE₂ and 5 μmol.L⁻¹ unlabelled PGE₂ in thepresence or absence of various concentrations of testing compounds inbinding buffer (50 mmol.L⁻¹ HEPES, pH 7.4, 5 mmol.L⁻¹ MgCl₂, 1 mmol.L⁻¹CaCl₂, 0.2% BSA) in a nonbinding 96-well plate, and incubated for 1-2 hat room temperature. Prior to filtration, a GF/C 96-well filter platewas coated with 0.33% polyethyleneimine for 30 min, then washed with 50mmol.L⁻¹ HEPES, pH 7.4, 0.5% BSA. Binding reactions were transferred tothe filter plate, and washed 3 times with Wash Buffer (1 mL per well perwash). The plate was dried and radioactivity counted. Binding of testingcompounds to other related prostanoid receptors was performed by MDSPharma Services (Bothell, Wash.) using a similar radiolabeled liganddisplacement method.

Exemplary compounds of the present invention were assayed according tothe methods set forth above in the radioligand EP4 receptor bindingassay described above. Table 3 below sets forth exemplary compounds ofthe present invention having Ki values as determined by the radioligandEP4 receptor binding assay described above.

TABLE 3 Ki Values of Exemplary Compounds EP4 binding Example StructureER-number Ki (μM) 103

ER-885290-00 0.014 104

ER-885740-00 0.043 105

ER-885741-00 0.016 106

ER-886045-00 0.050 107

ER-886046-00 0.008 108

ER-886074-00 0.013 109

ER-886078-00 0.008 110

ER-886090-00 0.010 111

ER-887480-00 0.026

Examples 112-113 In Vivo Biological Activity Example 112 Suppression ofArthritis Development in CIA Model

Male DBA/1 mice were immunized by injection at the base of the tail with0.1 mL emulsion containing 150 μg bovine type II collagen (bCII)emulsified in CFA. Three weeks after the 1^(st) immunization, all micewere boosted with bovine type II collagen emulsified in Freund'sincomplete adjuvant, ER-886046 was orally administered daily at a doseof 10, 30 or 100 mg.kg⁻¹ from day 20 after primary immunization butbefore disease onset (prophylactic evaluation) or after the diseaseinduction (therapeutic evaluation). The severity of arthritic symptomsin the paws of each mouse was graded every other days, double-blind,according to Williams R O (Collagen-induced arthritis as a model forrheumatoid arthritis. Methods Mol Med 2004, 98:207-216). Results aregiven in FIG. 1.

Example 113 Suppression of Arthritis Development in Glucose-6-PhosphateIsomerase (G6PI) Model

Male DBA/1 mice were immunized by injecting at the base of the tail 0.15mL of emulsion containing 300 μg recombinant humanGPI-glutathione-S-transferase (GST) fusion protein (hGPI) in CFA.ER-886046 was orally administered daily at a dose of 100 mg.kg⁻¹ fromday 6 after primary immunization but before disease onset (prophylacticevaluation) or after the disease induction (therapeutic evaluation).Each treatment group consisted of 6-8 mice. Arthritic animals wereclinically assessed every other day by an arthritis scoring system asfollows (Iwanami K, Matsumoto I, Tanaka-Watanabe Y, Mihira M, Ohsugi Y,Mamura M et al. Crucial role of IL-6/IL-17 axis in the induction ofarthritis by glucose-6-phosphate isomerase. Arthritis Rheum 2008,58:754-763): 0=no evidence of inflammation, 1=subtle inflammation orlocalized edema, 2=easily identified swelling but localized to dorsal orventral surface of paws, and score 3=swelling on all aspects of paws.Results are given in FIG. 2.

Certain Embodiments of the Invention

1. A compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl (e.g., monofluoromethyl, difluoromethyl, ortrifluoromethyl);

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy (e.g.,monofluoromethoxy, difluoromethoxy, or trifluoromethoxy);

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

2. The compound of embodiment 1, wherein

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; R₂ ismethyl, difluoromethyl, or trifluoromethyl;

R₃ is methyl;

R₄ is chloro, fluoro, trifluoromethyl, difluoromethyl, methyl, methoxy,difluoromethoxy, or trifluoromethoxy;

R₅ is hydrogen, chloro, fluoro, methyl, or methoxy;

R₆ and R₇ are hydrogen;

or a pharmaceutically acceptable salt thereof.

3. The compound of embodiment 2, wherein R₅ is hydrogen; or a

pharmaceutically acceptable salt thereof

4. The compound of embodiment 3, wherein R₄ is selected from chloro,trifluoromethyl, difluoromethyl, difluoromethoxy, and trifluoromethoxy;

or a pharmaceutically acceptable salt thereof.

5. The compound of any one of embodiments 1-4, wherein,

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl, and thecompound of Formula I consists of a mixture of stereoisomers;

or a pharmaceutically acceptable salt thereof.

6. The compound of any one of embodiments 1-4, wherein,

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl, and thecompound of Formula I consists of a substantially pure stereoisomer;

or a pharmaceutically acceptable salt thereof.

7. The compound of embodiment 6, wherein,

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl, and thecarbon of the compound of Formula I marked with a * has substantiallythe S-configuration;

or a pharmaceutically acceptable salt thereof.

8. The compound of embodiment 6, wherein,

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl, and thecarbon of the compound of Formula I marked with a * has substantiallythe R-configuration;

or a pharmaceutically acceptable salt thereof.

9. The compound of embodiment 1, wherein

R_(1a) and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl, trifluoromethyl, or difluoromethyl;

R₃ is methyl;

R₄ is trifluoromethyl, difluoromethyl, chloro, or fluoro;

R₆ and R₇ are hydrogen;

or a pharmaceutically acceptable salt thereof.

10. The compound of embodiment 1, selected from the group consisting of:

and pharmaceutically acceptable salts thereof.11. The compound of embodiment 10, which is:

or a pharmaceutically acceptable salt thereof.12. The compound of embodiment 10, which is:

or a pharmaceutically acceptable salt thereof.13. A pharmaceutical composition comprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or a pharmaceutically acceptable salt thereof;

and a pharmaceutically acceptable carrier.

14. A pharmaceutical composition comprising a compound of embodiment 10,or

a pharmaceutically acceptable salt thereof;

and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition comprising the compound which is:

or a pharmaceutically acceptable salt thereof;and a pharmaceutically acceptable carrier.16. A pharmaceutical composition comprising the compound which is:

or a pharmaceutically acceptable salt thereof;and a pharmaceutically acceptable carrier.17. A method of treating multiple sclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

18. A method of treating multiple sclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

19. A method of treating multiple sclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutical salt thereof.20. A method of treating multiple sclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutical salt thereof.21. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of multiple sclerosis.22. A method of treating rheumatoid arthritis in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

23. A method of treating rheumatoid arthritis in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising comprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

24. A method of treating rheumatoid arthritis in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.25. A method of treating rheumatoid arthritis in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.26. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of rheumatoid arthritis.27. A method of treating systemic lupus erythematosus in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

28. A method of treating systemic lupus erythematosus in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising a compound of embodiment 10,

or a pharmaceutically acceptable salt thereof.

29. A method of treating systemic lupus erythematosus in a mammal,comprising the step of administering to a mammal a pharmaceuticalcomposition comprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.30. A method of treating systemic lupus erythematosus in a mammal,comprising the step of administering to a mammal a pharmaceuticalcomposition comprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.31. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of systemic lupus erythematosus.32. A method of treating type 1 diabetes in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

33. A method of treating type 1 diabetes in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

34. A method of treating type 1 diabetes in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.35. A method of treating type 1 diabetes in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.36. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of type 1 diabetes.37. A method of treating psoriasis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising acompound of Formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

38. A method of treating psoriasis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising acompound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

39. A method of treating psoriasis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of Formula I which is:

or a pharmaceutically acceptable salt thereof.40. A method of treating psoriasis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of Formula I which is:

or a pharmaceutically acceptable salt thereof.41. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of psoriasis.42. A method of treating atherosclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of Formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

43. A method of treating atherosclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

44. A method of treating atherosclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof45. A method of treating atherosclerosis in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.46. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of atherosclerosis.47. A method of treating inflammatory pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of Formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

48. A method of treating inflammatory pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

49. A method of treating inflammatory pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.50. A method of treating inflammatory pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.51. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of inflammatory pain.52. A method of treating neuropathic pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of Formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R₁ andR_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

53. A method of treating neuropathic pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

54. A method of treating neuropathic pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.55. A method of treating neuropathic pain in a mammal, comprising thestep of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.56. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of neuropathic pain.57. A method of treating migraine-associated pain in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising a compound of Formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

58. A method of treating migraine-associated pain in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof

59. A method of treating migraine-associated pain in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.

60. A method of treating migraine-associated pain in a mammal,comprising the step of administering to the mammal a pharmaceuticalcomposition comprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.61. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of migraine-associated pain.62. A method of treating spondyloarthropathies in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising a compound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

63. A method of treating spondyloarthropathies in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

64. A method of treating spondyloarthropathies in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.65. A method of treating spondyloarthropathies in a mammal, comprisingthe step of administering to the mammal a pharmaceutical compositioncomprising the compound of Formula I which is:

or a pharmaceutically acceptable salt thereof.66. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of spondyloarthropathies.67. A method of treating cancer in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising acompound of formula I:

wherein:

one of R_(1a) and R_(1b) is hydrogen, and the other is methyl; or R_(1a)and R_(1b) are taken together to form a cyclopropyl ring;

R₂ is methyl or fluoromethyl;

R₃ is methyl;

R₄ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₅ is hydrogen, halo, fluoromethyl, methoxy, or fluoromethoxy;

R₆ is hydrogen, halo, methyl, or methoxy;

R₇ is hydrogen, halo, methyl, or methoxy; and

X is oxygen;

or pharmaceutically acceptable salts thereof.

68. A method of treating cancer in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprisingcomprising a compound of embodiment 10;

or a pharmaceutically acceptable salt thereof.

69. A method of treating cancer in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of Formula I which is:

or a pharmaceutically acceptable salt thereof.70. A method of treating cancer in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of Formula I which is:

or a pharmaceutically acceptable salt thereof.71. Use of a compound of embodiment 1 in the manufacture of a medicamentfor the treatment of cancer.72. The method of any of embodiments 67-71, wherein the cancer isselected from the group consisting of skin cancer, breast cancer,colorectal cancer, prostate cancer, kidney cancer, ovarian cancer,cervical cancer, endometrial cancer, glioblastoma, lung cancer, head andneck cancer, medulloblastoma, and urinary tract cancer.73. The method of embodiment 72, wherein the cancer is skin cancer.74. The method of embodiment 72, wherein the cancer is breast cancer.75. The method of embodiment 72, wherein the cancer is colorectalcancer.76. The method of embodiment 72, wherein the cancer is prostate cancer.77. The method of embodiment 72, wherein the cancer is kidney cancer.78. The method of embodiment 72, wherein the cancer is ovarian cancer.79. The method of embodiment 72, wherein the cancer is cervical cancer.80. The method of embodiment 72, wherein the cancer is endometrialcancer.81. The method of embodiment 72, wherein the cancer is glioblastoma.82. The method of embodiment 72, wherein the cancer is lung cancer.83. The method of embodiment 72, wherein the cancer is head and neckcancer.84. The method of embodiment 72, wherein the cancer is medulloblastoma.85. The method of embodiment 72, wherein the cancer is urinary tractcancer.

Other Embodiments

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example.

We claim:
 1. A compound of formula I:

wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl;or R_(1a) and R_(1b) are taken together to form a cyclopropyl ring; R₂is methyl or fluoromethyl; R₃ is methyl; R₄ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₅ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₆ is hydrogen, halo, methyl,or methoxy; R₇ is hydrogen, halo, methyl, or methoxy; and X is oxygen;or a pharmaceutically acceptable salt thereof.
 2. The compound of claim1, wherein: one of R_(1a) and R_(1b) is hydrogen, and the other ismethyl; R₂ is methyl, difluoromethyl, or trifluoromethyl; R₃ is methyl;R₄ is chloro, fluoro, trifluoromethyl, difluoromethyl, methyl, methoxy,difluoromethoxy, or trifluoromethoxy; R₅ is hydrogen, chloro, fluoro,methyl, or methoxy; and R₆ and R₇ are hydrogen; or a pharmaceuticallyacceptable salt thereof.
 3. The compound of claim 2, wherein R₅ ishydrogen; or a pharmaceutically acceptable salt thereof.
 4. The compoundof claim 3, wherein R₄ is selected from chloro, trifluoromethyl,difluoromethyl, difluoromethoxy, and trifluoromethoxy; or apharmaceutically acceptable salt thereof.
 5. The compound of claim 1,wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl,and the compound of Formula I is a mixture of stereoisomers; or apharmaceutically acceptable salt thereof.
 6. The compound of claim 1,wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl,and the compound of Formula I is a substantially pure stereoisomer; or apharmaceutically acceptable salt thereof.
 7. The compound of claim 6,wherein: the carbon of the compound of Formula I marked with an * hassubstantially the S-configuration; or a pharmaceutically acceptable saltthereof.
 8. The compound of claim 6, wherein: the carbon of the compoundof Formula I marked with an * has substantially the R-configuration; ora pharmaceutically acceptable salt thereof.
 9. The compound of claim 1,wherein R₁ and R_(1b) are taken together to form a cyclopropyl ring; R₂is methyl, trifluoromethyl, or difluoromethyl; R₃ is methyl; R₄ istrifluoromethyl, difluoromethyl, chloro, or fluoro; and R₆ and R₇ arehydrogen; or a pharmaceutically acceptable salt thereof.
 10. Thecompound of claim 1, wherein said compound is selected from the groupconsisting of:

and pharmaceutically acceptable salts thereof.
 11. The compound of claim1, wherein said compound is:

or a pharmaceutically acceptable salt thereof.
 12. The compound of claim1, wherein said compound is:

or a pharmaceutically acceptable salt thereof.
 13. A pharmaceuticalcomposition comprising a compound of formula I:

wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl;or R_(1a) and R_(1b) are taken together to form a cyclopropyl ring; R₂is methyl or fluoromethyl; R₃ is methyl; R₄ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₅ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₆ is hydrogen, halo, methyl,or methoxy; R₇ is hydrogen, halo, methyl, or methoxy; and X is oxygen;or a pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 14. A pharmaceutical composition comprising acompound of claim 10, or a pharmaceutically acceptable salt thereof; anda pharmaceutically acceptable carrier.
 15. A pharmaceutical compositioncomprising a compound:

or a pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 16. A pharmaceutical composition comprising acompound:

or a pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 17. A method of treating multiple sclerosis in amammal, comprising the step of administering to the mammal apharmaceutical composition comprising a compound of formula I:

wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl;or R_(1a) and R_(1b) are taken together to form a cyclopropyl ring; R₂is methyl or fluoromethyl; R₃ is methyl; R₄ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₅ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₆ is hydrogen, halo, methyl,or methoxy; R₇ is hydrogen, halo, methyl, or methoxy; and X is oxygen;or a pharmaceutically acceptable salt thereof.
 18. A method of treatingmultiple sclerosis in a mammal, comprising the step of administering tothe mammal a pharmaceutical composition comprising a compound of claim10, or a pharmaceutically acceptable salt thereof.
 19. A method oftreating multiple sclerosis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of claim 10 which is:

or a pharmaceutically acceptable salt thereof.
 20. A method of treatingmultiple sclerosis in a mammal, comprising the step of administering tothe mammal a pharmaceutical composition comprising the compound of claim10 which is:

or a pharmaceutically acceptable salt thereof.
 21. A method of treatingrheumatoid arthritis in a mammal, comprising the step of administeringto the mammal a pharmaceutical composition comprising a compound offormula I:

wherein: one of R_(1a) and R_(1b) is hydrogen, and the other is methyl;or R_(1a) and R_(1b) are taken together to form a cyclopropyl ring; R₂is methyl or fluoromethyl; R₃ is methyl; R₄ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₅ is hydrogen, halo, methyl,fluoromethyl, methoxy, or fluoromethoxy; R₆ is hydrogen, halo, methyl,or methoxy; R₇ is hydrogen, halo, methyl, or methoxy; and X is oxygen;or pharmaceutically acceptable salts thereof.
 22. A method of treatingrheumatoid arthritis in a mammal, comprising the step of administeringto the mammal a pharmaceutical composition comprising a compound ofclaim 10, or a pharmaceutically acceptable salt thereof.
 23. A method oftreating rheumatoid arthritis in a mammal, comprising the step ofadministering to the mammal a pharmaceutical composition comprising thecompound of claim 10 which is:

or a pharmaceutically acceptable salt thereof.
 24. A method of treatingrheumatoid arthritis in a mammal, comprising the step of administeringto the mammal a pharmaceutical composition comprising the compound ofclaim 10 which is:

or a pharmaceutically acceptable salt thereof.